title: Antimicrobial peptides isolated from Phyllomedusa nordestina (Amphibia) alter the permeability of plasma membrane of Leishmania and Trypanosoma cruzi.
abstract: Nature has provided inspiration for Drug Discovery studies and amphibian secretions have been used as a promising source of effective peptides which could be explored as novel drug prototypes for neglected parasitic diseases as Leishmaniasis and Chagas disease. In this study, we isolated four antimicrobial peptides (AMPs) from Phyllomedusa nordestina secretion, and studied their effectiveness against Leishmania (L.) infantum and Trypanosoma cruzi. The antiparasitic fractions were characterized by mass spectrometry and Edman degradation, leading to the identification of dermaseptins 1 and 4 and phylloseptins 7 and 8. T. cruzi trypomastigotes were susceptible to peptides, showing IC50 values in the range concentration of 0.25-0.68 μM. Leishmania (L.) infantum showed susceptibility to phylloseptin 7, presenting an IC50 value of 10 μM. Except for phylloseptin 7 which moderate showed cytotoxicity (IC50=34 μM), the peptides induced no cellular damage to mammalian cells. The lack of mitochondrial oxidative activity of parasites detected by the MTT assay, suggested that peptides were leishmanicidal and trypanocidal. By using the fluorescent probe SYTOX(®) Green, dermaseptins 1 and 4 and phylloseptins 7 and 8 showed time-dependent plasma membrane permeabilization of T. cruzi; phylloseptin 7 also showed a similar effect in Leishmania parasites. The present study demonstrates for the first time that AMPs target the plasma membrane of Leishmania and T. cruzi, leading to cellular death. Considering the potential of amphibian peptides against protozoan parasites and the reduced mammalian toxicity, they may contribute as scaffolds for drug design studies.
authors.affiliation: Departament of Parasitology, Instituto Adolfo Lutz, Av. Dr. Arnaldo 351, 8(o) Andar, 01246-000 São Paulo, SP, Brazil; Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, Av. Dr. Enéas de Carvalho Aguiar, 470, 05403-000 São Paulo, SP, Brazil.
authors.forename: Erika Gracielle
authors.initials: EG
authors.lastname: Pinto
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Pimenta
authors.forename: Marta Maria
authors.initials: MM
authors.lastname: Antoniazzi
authors.forename: Carlos
authors.initials: C
authors.lastname: Jared
authors.forename: Andre Gustavo
authors.initials: AG
authors.lastname: Tempone
mesh_terms: D029845:Amphibian Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D000981:Antiprotozoal Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001001:Anura / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D005260:Female; D005470:Fluorometry; D018314:Leishmania infantum / Q000187:drug effects* / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000187:drug effects; D013058:Mass Spectrometry; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D020539:Sequence Analysis, Protein; D014349:Trypanosoma cruzi / Q000187:drug effects* / Q000378:metabolism
chemical_list: D029845:Amphibian Proteins; D023181:Antimicrobial Cationic Peptides; D000981:Antiprotozoal Agents; C109653:dermaseptin 4 protein, Phyllomedusa; C585664:phylloseptin-7, Phyllomedusa; C585665:phylloseptin-8, Phyllomedusa; C070905:dermaseptin
keywords: Dermaseptin; Drugs; Leishmania; Peptides; Phylloseptin; Trypanosoma cruzi